Multiplex Assay Kit for Dickkopf Related Protein 1 (DKK1) ,etc. by FLIA (Flow Luminescence Immunoassay)

SK; Dickkopf Homolog 1(Xenopus Laevis)

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Dickkopf Related Protein 1 (DKK1) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Dickkopf Related Protein 1 (DKK1) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Dickkopf Related Protein 1 (DKK1) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Dickkopf Related Protein 1 (DKK1) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-94 81
EDTA plasma(n=5) 89-96 92
heparin plasma(n=5) 94-102 97
sodium citrate plasma(n=5) 86-93 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Dickkopf Related Protein 1 (DKK1) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Dickkopf Related Protein 1 (DKK1) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Dickkopf Related Protein 1 (DKK1) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 98-105% 87-104% 80-89% 97-105%
EDTA plasma(n=5) 82-94% 95-104% 92-99% 91-99%
heparin plasma(n=5) 92-101% 90-99% 97-105% 95-104%
sodium citrate plasma(n=5) 91-103% 93-101% 80-95% 97-104%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:DKK1) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Scientific Reports The possible role of Dickkopf-1, Golgi protein-73 and Midkine as predictors of hepatocarcinogenesis: a review and an Egyptian study Pubmed: 32198440
Nature Communications Immunological history governs human stem cell memory CD4 heterogeneity via the Wnt signaling pathway Pubmed: 32041953
JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY Circulating Osteocyte‐Related Biomarkers (vitamin D, sclerostin, dickkopf-1), hepcidin, and oxidative stress markers in early breast cancer: their impact in … Pubmed: 33065276
Oxid Med Cell Longev Long-Term Administration of Abacavir and Etravirine Impairs Semen Quality and Alters Redox System and Bone Metabolism in Growing Male Wistar Rats 34373766
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